哈萨克羊DQB2基因外显子2多态性及遗传变异分析

发布时间：2018-04-20 03:20

本文选题：DQB基因 + PCR-SSCP　；参考：《中国畜牧兽医》2017年12期

【摘要】：试验旨在研究DQB2基因外显子2多态性与哈萨克羊布鲁氏菌病易感性的相关性。通过PCR-SSCP技术对146只布鲁氏菌阴性哈萨克羊血液样本和28只布鲁氏菌阳性哈萨克羊血液样本中的白细胞表面抗原DQB2基因外显子2的多态性进行研究,挑取不同的等位基因进行克隆测序,经卡方检验分析每个SNP位点的基因频率、基因型频率及其多态性与布鲁氏菌病易感性的相关性,应用生物信息学软件分析与哈萨克羊布鲁氏菌病易感性相关的不同等位基因的mRNA二级结构及蛋白质的二级结构、三级结构和抗原表位。结果发现,在270bp的外显子2序列中共检测到33个SNPs,其中C9G、A180G位点的基因频率在病例组和对照组中的分布具有极显著性差异(P0.01),其基因型频率在病例组和对照组中存在显著差异(P0.05);A13T、C133G位点的基因频率在病例组和对照组中存在显著差异(P0.05);进一步分析发现,A180G突变位点的最小自由能最低,其mRNA二级结构最稳定;A13T和C133G2个突变位点均引起mRNA二级结构及蛋白质二级结构、三级结构和抗原表位的改变。本试验结果表明DQB2基因外显子2多态性与哈萨克羊布鲁氏菌病易感性呈显著相关。
[Abstract]:The aim of this study was to study the association between DQB2 gene exon 2 polymorphism and susceptibility to brucellosis in Kazak sheep. The polymorphisms of leukocyte surface antigen (DQB2) gene exon 2 in blood samples of 146 Brucella negative Kazakh sheep and 28 Brucella positive Kazakh sheep were studied by PCR-SSCP technique. Different alleles were selected for cloning and sequencing. The gene frequency, genotype frequency and polymorphism of each SNP locus were analyzed by chi-square test. Bioinformatics software was used to analyze the secondary structure of mRNA and the secondary structure of protein, tertiary structure and antigen epitope of different alleles associated with susceptibility to brucellosis in Kazak sheep. And it turns out, A total of 33 SNPs were detected in exon 2 of 270bp. There was a significant difference in the gene frequency of C9GnA180G locus between the case group and the control group (P0.01G). There was a significant difference in the genotype frequency between the case group and the control group (P0.05). There was significant difference in gene frequency between the two groups, and further analysis showed that the minimum free energy of A180G mutation was the lowest. The mutation sites of A13T and C133G2 in mRNA secondary structure caused the changes of secondary structure, protein secondary structure, tertiary structure and antigen epitope of mRNA. The results showed that the polymorphism of exon 2 of DQB2 gene was significantly associated with the susceptibility to brucellosis in Kazakh sheep.
【作者单位】：石河子大学生命科学学院;
【基金】：国家重点基础研究发展计划(973计划)(2010CB530200)
【分类号】：S826

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